Substituted pyrazolo[3,4-D]pyrimidines as p38 map kinase inhibitors

ABSTRACT

Compounds of the formula Ia or Ib: 
                         
wherein X and Y are N or one of X and Y is N and the other is CR g , and A, D, E, G, W, R 1 , R 2 , R 3 , R 4  and R 5  are as defined herein. Also disclosed are methods of making the subject compounds and methods of using the compounds for treatment of p38 MAP kinase-mediated diseases.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is entitled to the benefit of provisional patentapplication Ser. No. 60/712,010 filed on Aug. 25, 2005, the disclosureof which is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to fused pyrazolo pyrimidine derivativesand related compounds, a process for their manufacture, pharmaceuticalpreparations comprising the same, and methods for using the same.

BACKGROUND OF THE INVENTION

Mitogen-activated protein kinases (MAP) is a family of proline-directedserine/threonine kinases that activate their substrates by dualphosphorylation. The kinases are activated by a variety of signalsincluding nutritional and osmotic stress, UV light, growth factors,endotoxin and inflammatory cytokines. One group of MAP kinases is thep38 kinase group that includes various isoforms (e.g., p38α, p39β, p38γand p38δ). The p38 kinases are responsible for phosphorylating andactivating transcription factors as well as other kinases, and areactivated by physical and chemical stress, pro-inflammatory cytokinesand bacterial lipopolysaccharide.

More importantly, the products of the p38 phosphorylation have beenshown to mediate the production of inflammatory cytokines, including TNFand IL-1, and cyclooxygenase-2. Each of these cytokines has beenimplicated in numerous disease states and conditions. For example, TNF-αis a cytokine produced primarily by activated monocytes and macrophages.Its excessive or unregulated production has been implicated as playing acausative role in the pathogenesis of rheumatoid arthritis. Morerecently, inhibition of TNF production has been shown to have broadapplication in the treatment of inflammation, inflammatory boweldisease, multiple sclerosis and asthma.

TNF has also been implicated in viral infections, such as HIV, influenzavirus, and herpes virus including herpes simplex virus type-1 (HSV-1),herpes simplex virus type-2 (HSV-2), cytomegalovirus (CMV),varicella-zoster virus (VZV), Epstein-Barr virus, human herpes virus-6(HHV-6), human herpesvirus-7 (HHV-7), human herpesvirus-8 (HHV-8),pseudorabies and rhinotracheitis, among others.

Similarly, IL-1 is produced by activated monocytes and macrophages, andplays a role in many pathophysiological responses including rheumatoidarthritis, fever and reduction of bone resorption.

Additionally, the involvement of p38 has been implicated in stroke,Alzheimer's disease, osteoarthritis, lung injury, septic shock,angiogenesis, dermatitis, psoriasis and atopic dermatitis. J. Exp. Opin.Ther. Patents, 2000, 10(1).

The role of p38 MAP kinase as a therapeutic target in oncology has beenreviewed: Podar, K. H.; Teru; Chauhan, Dharminder; Anderson, Kenneth C.,“Targeting signalling pathways for the treatment of multiple myeloma”,Expert Opinion on therapeutic Targets 2005, 9, 359-381; Schultz, R. M.,“Potential of p38 MAP kinase inhibitors in the treatment of cancer”,Progress in Drug Research 2003, 60, 59-92.

The inhibition of these cytokines by inhibition of the p38 kinase is ofbenefit in controlling, reducing and alleviating many of these diseasestates.

SUMMARY

The invention provides compounds of formula Ia or 1b:

or pharmaceutically acceptable sals thereof,wherein:

n is from 0 to 4;

R¹ is optionally substituted phenyl;

R² is heteroaryl, heterocyclyl, —C(═O)—R^(a), —CN,—S(O)_(m)R^(a)—NR^(b)C(═O)—R^(a), —O—C(═O)—R^(a) or —NR^(b)SO₂R^(a),

wherein

-   -   m is from 0 to 2,    -   R^(a) is alkyl, heteroalkyl, amino, alkylamino, dialkylamino,        hydroxy or alkoxy, and    -   R^(b) is hydrogen or alkyl;

R³ is hydrogen or alkyl;

R⁴ is hydrogen, alkyl, hydroxy, amino, heteroalkyl, heteroalkoxy,heteroalkylamino, heterocyclyl, heterocyclylalkyl, hydroxycycloalkyl,cycloalkylalkyl, alkylsulfonyl, alkylsulfonamido, aryl, heteroaryl,aralkyl, heteroaralkyl, alkoxy, heteroaralkoxy,—(CHR^(c))_(r)—C(═O)—R^(d), —(CHR^(c))_(r)—O—C(═O)—R^(d),—(CHR^(c))_(r)—NH—C(═O)—R^(d) or —SO₂—R^(d),

wherein

-   -   R^(c) is hydrogen, alkyl or heteroalkyl;    -   R^(d) is alkyl, hydroxy, amino, heteroalkyl, aryl, aralkyl,        heteroaryl, or heterocyclyl;    -   r is from 0 to 4;

each R⁵ is independently alkyl, halo, haloalkyl, alkoxy, haloalkoxy,cyano, nitro, hydroxy, amino, heteroalkyl, heterocyclyl,heterocyclylalkyl, hydroxycycloalkyl, cycloalkylalkyl, alkoxyalkyloxy,aryl, heteroaryl, aralkyl, heteroaralkyl, aralkoxy, heteroaralkyoxy,cyanoalkoxy, alkenylalkoxy, alkynylalkoxy, hydroxyalkyloxy,hydroxyalkylalkynylalkoxy, —(CHR^(e))_(s)—C(═O)—R^(f),—(CHR^(e))_(s)—O—C(═O)—R^(f), —(CHR^(e))_(s)—NH—C(═O)—R^(f) or—SO₂—R^(f),

wherein

-   -   R^(e) is hydrogen, alkyl or heteroalkyl;    -   R^(f) is alkyl, hydroxy, amino, alkylamino, heteroalkyl, aryl,        aralkyl, heteroaryl, or heterocyclyl; and    -   s is from 0 to 4;

X and Y are nitrogen, or one of X and Y is nitrogen and the other isCR^(g);

wherein

-   -   R^(g) is hydrogen, alkyl, hydroxy, alkoxy, amino, haloalkyl,        cyano, halo, heteroalkyl, C(═O)—R^(h) or —SO₂—R^(h),

wherein

-   -   R^(h) is hydrogen or alkyl;

One or two of D, E and G is nitrogen, or D, E and G are carbon;

W is a bond, O, S(O)_(t), CH₂ or NR^(i); or W and R⁴ together formcyano;

wherein

-   -   t is from 0 to 2, and    -   R^(i) is hydrogen, alkyl, heteroalkyl, heterocyclyl,        hydroxycycloalkyl, —C(═O)—R^(j) or —SO₂—R^(j),    -   wherein        -   R^(j) is alkyl, aryl, aralkyl, heteroaryl, heteroalkyl or            heterocyclyl;    -   or R⁴ and R^(i) together with the atoms to which they are        attached may form a heterocyclic ring;

A is O, CH₂, S(O)_(u), C(═O), NR^(k), or CH(OR^(k)),

wherein

-   -   u is from 0 to 2, and    -   R^(k) is hydrogen or alkyl.

Another aspect of the invention provides a pharmaceutical formulationcomprising one or more compounds of formula I and a pharmaceuticallyacceptable carrier, diluent, and/or excipient therefore.

Compounds of the invention are inhibitors of protein kinases, andexhibit effective activity against p38 in vivo. They are selective forp38 kinase relative to cyclin-dependent kinases and tyrosine kinases.Therefore, compounds of the present invention can be used for thetreatment of diseases mediated by the pro-inflammatory cytokines such asTNF and IL-1. Thus, another aspect of the present invention provides amethod for treating p38 mediated diseases or conditions in which atherapeutically effective amount of one or more compounds of formula Iis administered to a patient.

DETAILED DESCRIPTION OF THE INVENTION

All publications cited in this disclosure are incorporated herein byreference in their entirety.

Definitions

Unless otherwise stated, the following terms used in this Application,including the specification and claims, have the definitions givenbelow. It must be noted that, as used in the specification and theappended claims, the singular forms “a”, “an,” and “the” include pluralreferents unless the context clearly dictates otherwise.

“Alkyl” means a linear saturated monovalent hydrocarbon moiety of one tosix carbon atoms or a branched saturated monovalent hydrocarbon moietyof three to six carbon atoms, e.g., methyl, ethyl, propyl, 2-propyl,n-butyl, iso-butyl, tert-butyl, pentyl, and the like.\

“Alkylene” means a linear saturated divalent hydrocarbon moiety of oneto six carbon atoms or a branched saturated divalent hydrocarbon moietyof three to six carbon atoms, e.g., methylene, ethylene,2,2-dimethylethylene, propylene, 2-methylpropylene, butylene, pentylene,and the like.

“Alkenyl” means a linear monovalent hydrocarbon radical of two to sixcarbon atoms or a branched monovalent hydrocarbon radical of three tosix carbon atoms, containing at least one double bond, e.g., ethenyl,propenyl, and the like. “Alkenlene means a divalent alkenyl radical.

“Alkynyl” means a linear monovalent hydrocarbon radical of two to sixcarbon atoms or a branched monovalent hydrocarbon radical of three tosix carbon atoms, containing at least one triple bond, e.g., ethynyl,propynyl, and the like. “Alkynylene” means a divalent alkynyl radical.

“Alkoxy” means a moiety of the formula —OR, wherein R is an alkyl moietyas defined herein. Examples of alkoxy moieties include, but are notlimited to, methoxy, ethoxy, isopropoxy, and the like.

“Alkoxyalkyl” means a moiety of the formula R^(a)—O—R^(b)—, where R^(a)is alkyl and R^(b) is alkylene as defined herein. Exemplary alkoxyalkylgroups include, by way of example, 2-methoxyethyl, 3-methoxypropyl,1-methyl-2-methoxyethyl, 1-(2-methoxyethyl)-3-methoxypropyl, and1-(2-methoxyethyl)-3-methoxypropyl.

“Alkoxyalkyloxy” and “alkoxyalkoxy”, which may be used interchangeably,mean a group of the formula —O—R—O—R′ wherein R′ is alkyl and R isalkylene as defined herein.

“Alkylamino means a moiety of the formula —NR—R′ wherein R is hyrdogenor alkyl and R′ is alkyl as defined herein.

“Alkoxyamino” means a moiety of the formula —NR—OR′ wherein R ishydrogen or alkyl and R′ is alkyl as defined herein.

“Alkylsulfanyl” means a moiety of the formula —SR wherein R is alkyl asdefined herein.

“Alkylsulfonyl” means a moiety of the formula —SO₂R wherein R is alkylas defined herein.

“Alkylsulfonylalkyl” means a moiety of the formula R^(a)—SO₂—R^(b)—,where R^(a) is alkyl and R^(b) is alkylene as defined herein. Exemplaryalkylsulfonylalkyl groups include, by way of example,3-methanesulfonylpropyl, 2-methanesulfonylethyl,2-methanesulfonylpropyl, and the like.

“Alkynylalkoxy” means a group of the formula —O—R—R′ wherein R isalkylene and R′ is alkynyl as defined herein.

“Amino” means a group —NR′R″ wherein R′ and R″ each independently ishydrogen or alkyl. “Amino” as used herein thus encompasses “alkylamino”and “dialkylamino”.

“Alkylaminoalkyl” means a group —R—NHR′ wherein R is alkylene and R′ isalkyl. Alkylaminoalkyl includes methylaminomethyl, methylaminoethyl,methylaminopropyl, ethylaminoethyl and the like.

“Dialkylaminoalkyl” means a group —R—NR′R″ wherein R is alkylene and R′and R″ are alkyl as defined herein. Dialkylaminoalkyl includesdimethylaminomethyl, dimethylaminoethyl, dimethylaminopropyl,N-methyl-N-ethylaminoethyl, and the like.

“Aminoalkoxy” means a group —OR—R′ wherein R′ is amino and R is alkyleneas defined herein.

“Alkylsulfonylamido” means a moiety of the formula —NR′SO₂—R wherein Ris alkyl and R′ is hydrogen or alkyl.

“Aryl” means a monovalent monocyclic or bicyclic aromatic hydrocarbonmoiety which is optionally substituted with one or more, preferably one,two or three, substituents, each of which is preferably selected fromthe group consisting of alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy,halo, nitro, cyano, amino, mono- and dialkylamino, methylenedioxy,ethylenedioxy, acyl, heteroalkyl, optionally substituted aryl,optionally substituted heteroaryl, optionally substituted aralkyl, andoptionally substituted heteroaralkyl. A particularly preferred arylsubstituent is halide. More specifically the term aryl includes, but isnot limited to, phenyl, 1-naphthyl, 2-naphthyl, and the like, each ofwhich can be substituted or unsubstituted.

“Aralkyl” refers to a moiety of the formula —R—R′ wherein R is alkyleneand R′ is aryl as defined herein.

“Aralkoxy” means a group —O—R—R′ wherein R is alkylene and R′ is aryl asdefined herein

“Substituted aralkyl” or “optionally substituted aralkyl” refers toaralkyl in which the aryl moiety is substituted or optionallysubstituted, respectively.

“Cyanoalkoxy” means a group of the formula —O—R—CN wherein R is alkyleneas defined herein.

“Cycloalkyl” refers to a saturated monovalent cyclic hydrocarbon moietyof three to seven ring carbons e.g., cyclopropyl, cyclobutyl,cyclohexyl, 4-methyl-cyclohexyl, and the like. Cycloalkyl may optionallybe substituted with one or more substituents, preferably one, two orthree, substituents. Preferably, cycloalkyl substituent is selected fromthe group consisting of alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy,halo, amino, mono- and dialkylamino, heteroalkyl, acyl, aryl andheteroaryl.

“Cycloalkylalkyl” refers to a moiety of the formula R^(c)—R^(d)—, whereR^(c) is cycloalkyl and R^(d) is alkylene as defined herein.

“Halo”, “halogen” and “halide” are used interchangeably herein and referto fluoro, chloro, bromo, or iodo. Preferred halides are fluoro andchloro with fluoro being a particularly preferred halide.

“Haloalkyl” means alkyl substituted with one or more same or differenthalo atoms, e.g., —CH₂Cl, —CF₃, —CH₂CF₃, —CH₂CCl₃, and the like.

“Heteroalkyl” means an alkyl moiety as defined herein wherein one ormore, preferably one, two or three, hydrogen atoms have been replacedwith a substituent independently selected from the group consisting of—OR^(a), —NR^(b)R^(c) (where n is 0 or 1 if R^(b) and R^(c) are bothindependently alkyl, cycloalkyl or cycloalkylalkyl, and 0 if not) and—S(O)_(n)R^(d) (where n is an integer from 0 to 2), with theunderstanding that the point of attachment of the heteroalkyl moiety isthrough a carbon atom, wherein R^(a) is hydrogen, acyl, alkoxycarbonyl,alkyl, hydroxyalkyl, alkoxyalkyl, alkylsulfonyl, aminocarbonyl,aminosulfonylamino, cycloalkyl, or cycloalkylalkyl; R^(b) and R^(c) areindependently of each other hydrogen, acyl, alkoxycarbonyl,aminocarbonyl, aminocarbonyl, aminosulfonylamino, hydroxyalkyl,alkoxyalkyl, alkylsulfonyl, cycloalkyl, cycloalkylalkyl, alkylsulfonyl,aminosulfonyl, mono- or di-alkylaminosulfonyl, aminoalkyl, mono- ordi-alkylaminoalkyl, hydroxyalkyl, alkoxyalkyl, hydroxyalkylsulfonyl oralkoxyalkylsulfonyl; and when n is 0, R^(d) is hydrogen, alkyl,cycloalkyl, cycloalkylalkyl, or aryl, and when n is 1 or 2, R^(d) isalkyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl, alkoxyalkyl,alkylamino, aminocarbonyl, aminosulfonylamino, alkylsulfonyl, amino, oroptionally substituted phenyl. Representative examples include, but arenot limited to, 2-hydroxyethyl, 3-hydroxypropyl,2-hydroxy-1-hydroxymethylethyl, 2,3-dihydroxypropyl,1-hydroxymethylethyl, 3-hydroxybutyl, 2,3-dihydroxybutyl,2-hydroxy-1-methylpropyl, 2-aminoethyl, 3-aminopropyl,2-methylsulfonylethyl, aminosulfonylmethyl, aminosulfonylethyl,aminosulfonylpropyl, methylaminosulfonylmethyl,methylaminosulfonylethyl, methylaminosulfonylpropyl, and the like.Accordingly, hydroxyalkyl and alkoxyalkyl are subset of heteroalkyl.

“Heteroaryl” means a monovalent monocyclic or bicyclic moiety of 5 to 12ring atoms having at least one aromatic ring containing one, two, orthree ring heteroatoms selected from N, O, or S (preferably N or O), theremaining ring atoms being C, with the understanding that the attachmentpoint of the heteroaryl moiety will be on an aromatic ring. Theheteroaryl ring is optionally substituted independently with one or moresubstituents, preferably one, two or three substituents, each of whichis independently selected from alkyl, haloalkyl, hydroxy, alkoxy, halo,nitro and cyano. More specifically the term heteroaryl includes, but isnot limited to, pyridyl, furanyl, thienyl, thiazolyl, isothiazolyl,triazolyl, imidazolyl, isoxazolyl, pyrrolyl, pyrazolyl, pyrimidinyl,benzofuranyl, tetrahydrobenzofuranyl, isobenzofuranyl, benzothiazolyl,benzoisothiazolyl, benzotriazolyl, indolyl, isoindolyl, benzoxazolyl,quinolyl, tetrahydroquinolinyl, isoquinolyl, benzimidazolyl,benzisoxazolyl or benzothienyl, imidazo[1,2-a]-pyridinyl,imidazo[2,1-b]thiazolyl, and the derivatives thereof.

“Heteroaralkyl” refers to a moiety of the formula —R—R′ wherein R isalkylene and R′ is heteroaryl as defined herein.

“Heteroaralkoxy” means a group —O—R—R′ wherein R is alkylene and R′ isheteroaryl as defined herein.

“Heterocyclyl” means a saturated or unsaturated non-aromatic cyclicmoiety of 3 to 8 ring atoms in which one or two ring atoms areheteroatoms selected from N, O, or S(O)_(n) (where n is an integer from0 to 2), preferably N or O, the remaining ring atoms being C, where oneor two C atoms may optionally be replaced by a carbonyl group. Theheterocyclyl ring may be optionally substituted independently with oneor more, preferably one, two, or three, substituents, each of which isindependently selected from alkyl, haloalkyl, hydroxyalkyl, halo, nitro,cyano, cyanoalkyl, hydroxy, alkoxy, amino, mono- and dialkylamino,aralkyl, —(X)_(n)—C(O)R^(e) (where X is O or NR^(f), n is 0 or 1, R^(e)is hydrogen, alkyl, haloalkyl, hydroxy (when n is 0), alkoxy, amino,mono- and dialkylamino, or optionally substituted phenyl, and R^(f) is Hor alkyl), -alkylene-C(O)R^(g) (where R^(g) is alkyl, —OR^(h) orNR^(i)R^(j) and R^(h) is hydrogen, alkyl or haloalkyl, and R^(i) andR^(j) are independently hydrogen or alkyl), and —S(O)_(n)R^(k) (where nis an integer from 0 to 2) such that when n is 0, R^(k) is hydrogen,alkyl, cycloalkyl, or cycloalkylalkyl, and when n is 1 or 2, R^(k) isalkyl, cycloalkyl, cycloalkylalkyl, amino, acylamino, monoalkylamino, ordialkylamino. A particularly preferred group of heterocyclylsubstituents include alkyl, haloalkyl, hydroxyalkyl, halo, hydroxy,alkoxy, amino, mono- and dialkylamino, aralkyl, and —S(O)_(n)R^(k). Inparticular, the term heterocyclyl includes, but is not limited to,tetrahydrofuranyl, pyridinyl, tetrahydropyranyl, piperidino,N-methylpiperidin-3-yl, piperazino, N-methylpyrrolidin-3-yl,3-pyrrolidino, morpholino, thiomorpholino, thiomorpholino-1-oxide,thiomorpholino-1,1-dioxide, 4-(1,1-dioxo-tetrahydro-2H-thiopyranyl),pyrrolinyl, imidazolinyl, N-methanesulfonyl-piperidin-4-yl, and thederivatives thereof, each of which may be optionally substituted.

“Heterocyclylalkyl” means a moiety of the formula —R—R′ wherein R isalkylene and R′ is heterocyclyl as defined herein.

“Heterocyclyloxy” means a moiety of the formula —OR wherein R isheterocyclyl as defined herein.

“Heterocyclylalkoxy” means a moiety of the formula —OR—R′ wherein R isalkylene and R′ is heterocyclyl as defined herein.

“Hydroxyalkoxy” means a moiety of the formula —OR wherein R ishydroxyalkyl as defined herein.

“Hydroxyalkylamino” means a moiety of the formula —NR—R′ wherein R ishydrogen or alkyl and R′ is hydroxyalkyl as defined herein.

“Hydroxyalkylaminoalkyl” means a moiety of the formula —R—NR′—R″ whereinR is alkylene, R′ is hydrogen or alkyl, and R″ is hydroxyalkyl asdefined herein.

“Hydroxyalkyl” refers to a subset of heteroalkyl and refers inparticular to an alkyl moiety as defined herein that is substituted withone or more, preferably one, two or three hydroxy groups, provided thatthe same carbon atom does not carry more than one hydroxy group.Representative examples include, but are not limited to, hydroxymethyl,2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl,4-hydroxybutyl, 2,3-dihydroxypropyl, 2-hydroxy-1-hydroxymethylethyl,2,3-dihydroxybutyl, 3,4-dihydroxybutyl and2-(hydroxymethyl)-3-hydroxypropyl

“Hydroxycycloalkyl” refers to a subset of cycloalkyl moiety as definedherein and specifically refers to a cycloalkyl moiety as defined hereinwhere one or more, preferably one, two or three, hydrogen atoms in thecycloalkyl moiety have been replaced with a hydroxy substituent.Representative examples include, but are not limited to, 2-, 3-, or4-hydroxycyclohexyl, and the like.

“Hydroxyalkyloxy” means a group of the formula —O—R—OH wherein R isalkylene as defined herein.

“Hydroxyalkylalkynylalkoxy” means a group of the formula —O—R—R′—R″—OHwherein R and R″ are alkylene and R′ is alkynylene as defined herein.

“Leaving group” has the meaning conventionally associated with it insynthetic organic chemistry, i.e., an atom or a group capable of beingdisplaced by a nucleophile and includes halo (such as chloro, bromo, andiodo), alkanesulfonyloxy, arenesulfonyloxy, alkylcarbonyloxy (e.g.,acetoxy), arylcarbonyloxy, mesyloxy, tosyloxy,trifluoromethanesulfonyloxy, aryloxy (e.g., 2,4-dinitrophenoxy),methoxy, N,O-dimethylhydroxylamino, and the like.

“Optionally substituted”, when used in association with “aryl”, phenyl”,“heteroaryl” “cycloalkyl” or “heterocyclyl”, means an aryl, phenyl,heteroaryl, cycloalkylyl or heterocyclyl which is optionally substitutedindependently with one to four substituents, preferably one or twosubstituents selected from alkyl, cycloalkyl, cycloalkylalkyl,heteroalkyl, hydroxyalkyl, halo, nitro, cyano, hydroxy, alkoxy, amino,acylamino, mono-alkylamino, di-alkylamino, haloalkyl, haloalkoxy,heteroalkyl, —COR (where R is hydrogen, alkyl, phenyl or phenylalkyl),—(CR′R″)_(n)—COOR (where n is an integer from 0 to 5, R′ and R″ areindependently hydrogen or alkyl, and R is hydrogen, alkyl, cycloalkyl,cycloalkylalkyl, phenyl or phenylalkyl), or —(CR′R″)_(n)—CONR^(a)R^(b)(where n is an integer from 0 to 5, R′ and R″ are independently hydrogenor alkyl, and R^(a) and R^(b) are, independently of each other,hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl or phenylalkyl), oras provided herein elsewhere.

“Pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition that is generally safe,non-toxic and neither biologically nor otherwise undesirable, andincludes excipient that is acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the specification and claims includes both one and more than onesuch excipient.

“Pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like.

“Protecting group” refers to a grouping of atoms that when attached to areactive group in a molecule masks, reduces or prevents that reactivity.Examples of protecting groups can be found in T. W. Green and P. G.Futs, Protective Groups in Organic Chemistry, (Wiley, 2^(nd) ed. 1991)and Harrison and Harrison et al., Compendium of Synthetic OrganicMethods, Vols. 1-8 (John Wiley and Sons, 1971-1996). Representativeamino protecting groups include, formyl, acetyl, trifluoroacetyl,benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl (Boc), trimethylsilyl (TMS), 2-trimethylsilyl-ethanesulfonyl (SES), trityl andsubstituted trityl groups, allyloxycarbonyl,9-fluorenylmethyloxycarbonyl (FMOC), nitro-veratryloxycarbonyl (NVOC),and the like. Representative hydroxy protecting groups include thosewhere the hydroxy group is either acylated or alkylated such as benzyl,and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers,trialkylsilyl ethers and allyl ethers.

“Treating” or “treatment” of a disease includes: (1) preventing thedisease, i.e., causing the clinical symptoms of the disease not todevelop in a mammal that may be exposed to or predisposed to the diseasebut does not yet experience or display symptoms of the disease; (2)inhibiting the disease, i.e., arresting or reducing the development ofthe disease or its clinical symptoms; or (3) relieving the disease,i.e., causing regression of the disease or its clinical symptoms.

“A therapeutically effective amount” means the amount of a compoundthat, when administered to a mammal for treating a disease, issufficient to effect such treatment for the disease. The“therapeutically effective amount” will vary depending on the compound,the disease and its severity and the age, weight, etc., of the mammal tobe treated.

As used herein, the terms “those defined above” and “those definedherein” are used interchangeably herein and, when referring to avariable, incorporates by reference the broad definition of the variableas well as preferred, more preferred and most preferred definitions, ifany.

“Modulator” means a molecule that interacts with a target. Theinteractions include, but are not limited to, agonist, antagonist, andthe like, as defined herein.

“Optional” or “optionally” means that the subsequently described eventor circumstance may but need not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not.

“Disease state” means any disease, condition, symptom, or indication.

“Inert organic solvent” or “inert solvent” means the solvent is inertunder the conditions of the reaction being described in conjunctiontherewith, including for example, benzene, toluene, acetonitrile,tetrahydrofuran, N,N-dimethylformamide, chloroform, methylene chlorideor dichloromethane, dichloroethane, diethyl ether, ethyl acetate,acetone, methyl ethyl ketone, methanol, ethanol, propanol, isopropanol,tert-butanol, dioxane, pyridine, and the like. Unless specified to thecontrary, the solvents used in the reactions of the present inventionare inert solvents.

“Solvates” means solvent addition forms that contain eitherstoichiometric or non stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate, when the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one of the substances in whichthe water retains its molecular state as H₂O, such combination beingable to form one or more hydrate.

“Subject” means mammals and non-mammals. Mammals means any member of themammalia class including, but not limited to, humans; non-human primatessuch as chimpanzees and other apes and monkey species; farm animals suchas cattle, horses, sheep, goats, and swine; domestic animals such asrabbits, dogs, and cats; laboratory animals including rodents, such asrats, mice, and guinea pigs; and the like. Examples of non-mammalsinclude, but are not limited to, birds, and the like. The term “subject”does not denote a particular age or sex.

The terms “those defined above” and “those defined herein” whenreferring to a variable incorporates by reference the broad definitionof the variable as well as preferred, more preferred and most preferreddefinitions, if any.

The terms “treating”, “contacting” and “reacting” when referring to achemical reaction means adding or mixing two or more reagents underappropriate conditions to produce the indicated and/or the desiredproduct. It should be appreciated that the reaction which produces theindicated and/or the desired product may not necessarily result directlyfrom the combination of two reagents which were initially added, i.e.,there may be one or more intermediates which are produced in the mixturewhich ultimately leads to the formation of the indicated and/or thedesired product.

Nomenclature

In general, the nomenclature used in this Application is based onAUTONOM™ v.4.0, a Beilstein Institute computerized system for thegeneration of IUPAC systematic nomenclature. Chemical structures shownherein were prepared using ISIS® version 2.2. Any open valency appearingon a carbon, oxygen or nitrogen atom in the structures herein indicatesthe presence of a hydrogen. Where a chiral center is present in astructure but no specific enantiomer is shown, the structure encompassesboth enantiomers associated with the chiral center.

Compounds of the Invention

The invention provides compounds of formula Ia or Ib:

or pharmaceutically acceptable sals thereof,wherein:

n is from 0 to 4;

R¹ is optionally substituted phenyl;

-   R² is heteroaryl, heterocyclyl, —C(═O)—R^(a), —CN,    —S(O)_(m)R^(a)—NR^(b)C(═O)—R^(a), —O—C(═O)—R^(a) or —NR^(b)SO₂R^(a),

wherein

-   -   m is from 0 to 2,    -   R^(a) is alkyl, heteroalkyl, amino, alkylamino, dialkylamino,        hydroxy or alkoxy, and    -   R^(b) is hydrogen or alkyl;

R³ is hydrogen or alkyl;

R⁴ is hydrogen, alkyl, hydroxy, amino, heteroalkyl, heteroalkoxy,heteroalkylamino, heterocyclyl, heterocyclylalkyl, hydroxycycloalkyl,cycloalkylalkyl, alkylsulfonyl, alkylsulfonamido, aryl, heteroaryl,aralkyl, heteroaralkyl, alkoxy, heteroaralkoxy,—(CHR^(c))_(r)—C(═O)—R^(d), —(CHR^(c))_(r)—O—C(═O)—R^(d),—(CHR^(c))_(r)—NH—C(═O)—R^(d) or —SO₂—R^(d),

wherein

-   -   R^(c) is hydrogen, alkyl or heteroalkyl;    -   R^(d) is alkyl, hydroxy, amino, heteroalkyl, aryl, aralkyl,        heteroaryl, or heterocyclyl;    -   r is from 0 to 4;

each R⁵ is independently alkyl, halo, haloalkyl, alkoxy, haloalkoxy,cyano, nitro, hydroxy, amino, heteroalkyl, heterocyclyl,heterocyclylalkyl, hydroxycycloalkyl, cycloalkylalkyl, alkoxyalkyloxy,aryl, heteroaryl, aralkyl, heteroaralkyl, aralkoxy, heteroaralkyoxy,cyanoalkoxy, alkenylalkoxy, alkynylalkoxy, hydroxyalkyloxy,hydroxyalkylalkynylalkoxy, —(CHR^(e))_(s)—C(═O)—R^(f),—(CHR^(e))_(s)—O—C(═O)—R^(f), —(CHR^(e))_(s)—NH—C(═O)—R^(f) or—SO₂—R^(f),

wherein

-   -   R^(e) is hydrogen, alkyl or heteroalkyl;    -   R^(f) is alkyl, hydroxy, amino, alkylamino, heteroalkyl, aryl,        aralkyl, heteroaryl, or heterocyclyl; and    -   s is from 0 to 4;

X and Y are nitrogen, or one of X and Y is nitrogen and the other isCR^(g);

wherein

-   -   R^(g) is hydrogen, alkyl, hydroxy, alkoxy, amino, haloalkyl,        cyano, halo, heteroalkyl, C(═O)R^(h) or —SO₂—R^(h),    -   wherein        -   R^(h) is hydrogen or alkyl;

One or two of D, E and G is nitrogen, or D, E and G are carbon;

W is a bond, O, S(O)_(t), CH₂ or NR^(i); or W and R⁴ together formcyano;

wherein

-   -   t is from 0 to 2, and    -   R^(i) is hydrogen, alkyl, heteroalkyl, heterocyclyl,        hydroxycycloalkyl,    -   —C(═O)—R^(j) or —SO₂—R^(j),    -   wherein        -   R^(j) is alkyl, aryl, aralkyl, heteroaryl, heteroalkyl or            heterocyclyl;    -   or R⁴ and R^(i) together with the atoms to which they are        attached may form a heterocyclic ring;

A is O, CH₂, S(O)_(u), C(═O), NR^(k), or CH(OR^(k)),

wherein

-   -   u is from 0 to 2, and    -   R^(k) is hydrogen or alkyl.

In many embodiments, the compounds of the invention are of formula Ia.

In certain embodiments of formula Ia or formula 1b, R³ is hydrogen.

In certain embodiments of formula Ia or formula 1b, D is nitrogen.

In certain embodiments of formula Ia or formula 1b, E is nitrogen.

In certain embodiments of formula Ia or formula 1b, G is nitrogen.

In certain embodiments of formula Ia or formula 1b, D and G arenitrogen.

In certain embodiments of formula Ia or formula 1b, D, E and G arecarbon.

In certain embodiments of formula Ia or formula 1b, D and E are carbonand G is nitrogen.

In certain embodiments of formula Ia or formula 1b, X and Y arenitrogen. In other embodiments of formula Ia, on of X and Y is nitrogenand the other is CR^(e).

In certain embodiments of formula Ia or formula 1b, X is nitrogen and Yis CR^(e).

In certain embodiments of formula Ia or formula 1b, X is CR^(e) and Y isnitrogen.

In certain embodiments of formula Ia or formula 1b, A is O, S or NR^(i).Preferably in such embodiments A is O.

In certain embodiments of formula Ia or formula 1b, R¹ is 2-halophenylor 2,4-dihalophenyl.

In certain embodiments of formula Ia or formula 1b, W is NR^(f) or O andR⁴ is heteroalkyl.

In certain embodiments of formula Ia or formula 1b, W is NR^(f) or O andR⁴ is hydroxyalkyl, alkoxyalkyl or alkylsulfonylalkyl.

In certain embodiments of formula Ia or formula 1b, W is NR^(f) or O andR⁴ is hydroxyalkyl.

In certain embodiments of formula Ia or formula 1b, W is a bond and R⁴is hydrogen.

In certain embodiments of formula Ia or formula 1b, R² is—S(O)_(m)R^(a).

In certain embodiments of formula Ia or formula 1b, R² is —S(O)_(m)R^(a)and m is 2.

In certain embodiments of formula Ia or formula 1b, R² is —S(O)_(m)R^(a)and m is 0.

In embodiments of formula Ia wherein R^(a) is heteroalkyl, suchheteroalkyl is preferably hydroxyalkyl, aminoalkyl, alkoxyalkyl,alkylsulfonylalkyl, alkylsulfonamidoalkyl or alkylaminosulonamidoalkyl.More preferably such heteralkyl is hydroxyalkyl, aminoalkyl, alkoxyalkylor alkylsulfonylalkyl. Still more preferably such heteroalkyl ishydroxyalkyl or alkoxyalkyl.

In certain embodiments of formula Ia or formula 1b, n is 1, R⁵ is at the2-position of the phenyl ring to which it is attached, and R² is—S(O)_(m)—R^(a) at the 5-position of the phenyl ring to which it isattached.

In certain embodiments of formula Ia or formula 1b, R² is a five or sixmembered monocyclic heteroaryl containing one or two heteroatomsselected from O, N and S.

In certain embodiments of formula Ia or formula 1b, R² is thienyl,furanyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl,isoxazolyl or isothiazolyl.

In certain embodiments of formula Ia or formula 1b, R² is pyridyl,pyrimidinyl or pyrazolyl.

In certain embodiments of formula Ia or formula 1b, R² is a five or sixmembered monocyclic heterocyclyl containing one or two heteroatomsselected from O, N and S.

In certain embodiments of formula Ia or formula 1b, R² istetrahydrofuranyl, morpholinyl or pyrrolyl.

In certain embodiments of formula Ia or formula 1b, R² is piperidinyl,piperazinyl, morpholinyl or tetrahydropyranyl, preferablymorpholin-4-yl.

In certain embodiments of formula Ia, R² is —C(═O)—R^(a).

In certain embodiments of formula Ia, R² is —NR^(b)SO₂R^(a).

In certain embodiments of formula Ia or formula 1b, R² is —CN.

In certain embodiments of formula Ia or formula 1b, D, E and G arecarbon and R³ is hydrogen.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen and X and Y are nitrogen.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, R² is —S(O)_(m)—R^(a), R^(a) is alkyl orhydroxyalkyl, and m is 2.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, R² is —S(O)_(m)—R^(a), R^(a) is alkyl orhydroxyalkyl, m is 2, and A is O.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, R² is —S(O)_(m)—R^(a), R^(a) is alkyl orhydroxyalkyl, m is 2, A is O, and R¹ is 2-halophenyl or2,4-dihalophenyl.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, R² is —S(O)_(m)—R^(a), R^(a) is alkyl orhydroxyalkyl, m is 2, A is O, R¹ is 2-halophenyl or 2,4-dihalophenyl,and n is 0 or 1.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, R² is —S(O)_(m)—R^(a), R^(a) is alkyl orhydroxyalkyl, m is 2, A is O, R¹ is 2-halophenyl or 2,4-dihalophenyl, nis 0 or 1, and R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, R² is —S(O)_(m)—R^(a), R^(a) is alkyl orhydroxyalkyl, m is 2, A is O, R¹ is 2-halophenyl or 2,4-dihalophenyl, nis 0 or 1, R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, andR^(a) is alkyl.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, R² is —S(O)_(m)—R^(a), R^(a) is alkyl orhydroxyalkyl, m is 2, A is O, R¹ is 2-halophenyl or 2,4-dihalophenyl, nis 0 or 1, R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, R^(a)is alkyl, and W is NR^(f) or O and R⁴ is heteroalkyl. Preferredheteroalkyl in such embodiments include hydroxyalkyl, aminoalkyl,alkoxyalkyl, alkylsulfonylalkyl, alkylsulfonamidoalkyl andalkylaminosulonamidoalkyl. More preferably such heteralkyl ishydroxyalkyl, aminoalkyl, alkoxyalkyl or alkylsulfonylalkyl.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, R² is —S(O)_(m)—R^(a), m is 2, A is O,R¹ is 2-halophenyl or 2,4-dihalophenyl, n is 0 or 1, R⁵ is alkyl,alkoxy, halo, alkoxyalkoxy or benzyloxy, R^(a) is alkyl or hydroxyalkyl,W is a bond and R⁴ is hydrogen.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, m is 2, A is O, R¹ is 2-halophenyl or2,4-dihalophenyl, n is 0 or 1, R⁵ is alkyl, alkoxy, halo, alkoxyalkoxyor benzyloxy located at the 2-position of the phenyl ring to which it isattached, R² is —S(O)_(m)—R^(a) located at the 5-position of the phenylring to which it is attached, m is 2 and R^(a) is alkyl or hydroxyalkyl.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, m is 2, A is O, R¹ is 2-halophenyl or2,4-dihalophenyl, n is 1, R⁵ is halo located at the 2-position of thephenyl ring to which it is attached, R² is —S(O)_(m)—R^(a) located atthe 5-position of the phenyl ring to which it is attached, m is 2 andR^(a) is alkyl or hydroxyalkyl.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, m is 2, A is O, R¹ is 2-halophenyl or2,4-dihalophenyl, n is 1, R⁵ is halo located at the 2-position of thephenyl ring to which it is attached, R² is —S(O)_(m)—R^(a) located atthe 5-position of the phenyl ring to which it is attached, m is 2, R^(a)is alkyl, W is a bond, and R⁴ is hydrogen.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, m is 2, A is O, R¹ is 2-halophenyl or2,4-dihalophenyl, n is 1, R⁵ is halo located at the 2-position of thephenyl ring to which it is attached, R² is —S(O)_(m)—R^(a) located atthe 5-position of the phenyl ring to which it is attached, m is 2, R^(a)is alkyl, W is O or NR^(f), and R⁴ is hydroxyalkyl, alkoxyalkyl oralkylsulfonyl alkyl, preferably hydroxyalkyl.

In certain embodiments of formula Ia, D and E are carbon, G is nitrogen,R³ is hydrogen, X and Y are nitrogen, m is 2, A is O, R¹ is 2-halophenylor 2,4-dihalophenyl, n is 1, R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy orbenzyloxy located at the 2-position of the phenyl ring to which it isattached, R² is —S(O)_(m)—R^(a) located at the 5-position of the phenylring to which it is attached, m is 2 and R^(a) is alkyl or hydroxyalkyl.

In certain embodiments of formula Ia, D, E and G are carbon, R³ ishydrogen, X and Y are nitrogen, m is 2, A is O, R¹ is 2-halophenyl or2,4-dihalophenyl, n is 1, R⁵ is halo located at the 2-position of thephenyl ring to which it is attached, R² is —S(O)_(m)—R^(a) located atthe 5-position of the phenyl ring to which it is attached, m is 2 andR^(a) is methyl.

In certain embodiments of formula Ia, D and E are carbon, G is nitrogen,R³ is hydrogen, X and Y are nitrogen, m is 2, A is O, R¹ is 2-halophenylor 2,4-dihalophenyl, n is 1, R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy orbenzyloxy located at the 2-position of the phenyl ring to which it isattached, R² is —S(O)_(m)—R^(a) located at the 5-position of the phenylring to which it is attached, m is 2 and R^(a) is alkyl.

In certain embodiments of formula Ia, the subject compounds are morespecifically of the formula:

wherein:

p is from 0 to 4;

each R⁶ is independently halo, alkyl, alkoxy, haloalkyl, haloalkoxy orcyano; and

n, W, R^(a), R⁴ and R⁵ are as defined herein.

In certain embodiments of formula II, W is NR^(f) or O and R⁴ isheteroalkyl.

In certain embodiments of formula II, W is a bond and R⁴ is hydrogen.

In certain embodiments of formula II, p is 1 or 2 and R⁶ is halo,preferably fluoro.

In certain embodiments of formula II, n is 0 or 1, and R⁵ is alkyl,alkoxy, halo, alkoxyalkoxy or benzyloxy.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 0 or1, and R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 0 or1, and R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, and R^(a)is alkyl or hydroxyalkyl.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 0 or1, and R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, and R^(a)is heteroalkyl, preferably hydroxyalkyl, alkoxyalkyl oralkylsulfonylalkyl.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 0 or1, R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, R^(a) is alkylor hydroxyalkyl, and m is 2.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 0 or1, R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy located at the2-position of the phenyl to which it is attached, R^(a) is alkyl orhydroxyalkyl, m is 2, and the group R^(a)—S(O)_(m)— is located at the5-position of the phenyl ring to which it is attached.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 0 or1, and R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, R^(a) isalkyl or hydroxyalkyl, W is NR^(f) or O and R⁴ is heteroalkyl. Preferredheteroalkyl in such embodiments include hydroxyalkyl, aminoalkyl,alkoxyalkyl, alkylsulfonylalkyl, alkylsulfonamidoalkyl andalkylaminosulonamidoalkyl. More preferably such heteroalkyl ishydroxyalkyl, aminoalkyl, alkoxyalkyl or alkylsulfonylalkyl.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 0 or1, and R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, R² isalkyl, W is a bond and R⁴ is hydrogen.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 1,R⁵ is halo, R^(a) is alkyl or hydroxyalkyl, W is NR^(f) or O, and R⁴ ishydroxyalkyl, alkoxyalkyl or alkylsulfonylalkyl.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 1,R⁵ is halo, R^(a) is alkyl or hydroxyalkyl, W is a bond, and R⁴ ishydrogen.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 1,R⁵ is halo, R^(a) is methyl, W is NR^(f) or O, and R⁴ is hydroxyalkyl,alkoxyalkyl or alkylsulfonylalkyl, preferably hydroxyalkyl.

In certain embodiments of formula II, p is 1 or 2, R⁶ is halo, n is 1,R⁵ is halo, R^(a) is methyl, W is a bond, and R⁴ is hydrogen.

In certain embodiments of formula Ia, the subject compounds are morespecifically of the formula:

wherein W, R², R⁴ and R⁵ are as defined herein.

In certain embodiments of formula III, R^(a) is alkyl or hydroxyalkyl.

In certain embodiments of formula III, R^(a) is heteroalkyl, preferablyhydroxyalkyl, alkoxyalkyl or alkylsulfonylalkyl, and still morepreferably hydroxyalkyl.

In certain embodiments of formula III, W is NR^(f) or O and R⁴ isheteroalkyl. Preferred heteroalkyl in such embodiments includehydroxyalkyl, aminoalkyl, alkoxyalkyl, alkylsulfonylalkyl,alkylsulfonamidoalkyl and alkylaminosulonamidoalkyl. More preferablysuch heteralkyl is hydroxyalkyl, aminoalkyl, alkoxyalkyl oralkylsulfonylalkyl.

In certain embodiments of formula III, W is a bond and R⁴ is hydrogen.

In certain embodiments of formula III, R⁵ is alkyl, alkoxy, halo,alkoxyalkoxy or benzyloxy.

In certain embodiments of formula III, R^(a) is alkyl and R⁵ is alkyl,alkoxy, halo, alkoxyalkoxy or benzyloxy.

In certain embodiments of formula III, R^(a) is alkyl or hydroxyalkyl,R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, W is NR^(f) or Oand R⁴ is heteroalkyl.

In certain embodiments of formula III, R^(a) is alkyl or hydroxyalkyl,R⁵ is alkyl, alkoxy, halo, alkoxyalkoxy or benzyloxy, W is a bond and R⁴is hydrogen.

In certain embodiments of formula III, R^(a) is alkyl, R⁵ is halo,alkoxyalkoxy or benzyloxy, W is NR^(f) or O, and R⁴ is hydroxyalkyl,alkoxyalkyl or alkylsulfonylalkyl.

In certain embodiments of formula III, R^(a) is alkyl, R⁵ is halo,alkoxyalkoxy or benzyloxy, W is a bond, and R⁴ is hydrogen.

In certain embodiments of formula III, R^(a) is methyl, R⁵ is halo, W isNR^(f) or O, and R⁴ is hydroxyalkyl, alkoxyalkyl or alkylsulfonylalkyl.

In certain embodiments of formula III, R^(a) is methyl, R⁵ is halo, W isa bond and R⁴ is hydrogen.

In embodiments of the invention where any of R¹, R², R³, R⁴, R⁵, R⁶,R^(a), R^(b), R^(c), R^(d), R^(e), R^(f), R^(g), R^(h), R^(i), R^(j) orR^(k) is alkyl or contains an alkyl moiety, such alkyl is preferablylower alkyl, i.e. C₁-C₆alkyl, and more preferably C₁-C₄alkyl.

Pharmaceutically acceptable acid addition salts of the compounds ofFormula I include salts derived from inorganic acids such ashydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic,phosphorous, and the like, as well as the salts derived from organicacids, such as aliphatic mono- and dicarboxylic acids,phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioicacids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Suchsalts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite,nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate,metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate,propionate, caprylate, isobutyrate, oxalate, malonate, succinate,suberate, sebacate, fumarate, maleate, mandelate, benzoate,chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate,benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate,maleate, tartrate, methanesulfonate, and the like. Also contemplated aresalts of amino acids such as arginate and the like and gluconate,galacturonate (see, for example, Berge S. M., et al., “PharmaceuticalSalts,” J. of pharmaceutical Science, 1977, 66, 1-19).

The acid addition salts of the basic compounds can be prepared bycontacting the free base form with a sufficient amount of the desiredacid to produce the salt in the conventional manner. The free base formcan be regenerated by contacting the salt form with a base and isolatingthe free base in the conventional manner. The free base forms differfrom their respective salt forms somewhat in certain physical propertiessuch as solubility in polar solvents, but otherwise the salts areequivalent to their respective free base for purposes of the presentinvention.

Representative compounds in accordance with the invention are shownbelow in Table 1.

TABLE 1 # Structure Name MP/M + H 1

6-(2,4-Difluoro-phenoxy)-3- (3-methanesulfonyl-phenyl)- 1H-pyrazolo[3,4-d]pyrimidine 403 2

3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine 216.5-218.2° C. 3

6-(2,4-Difluoro-phenoxy)-3- (5-methanesulfonyl-2- methoxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidine 433 4

2-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- ylamino]-propane-1,3-diol 527 5

6-(2,4-Difluoro-phenoxy)-3- (5-methanesulfonyl-2-methyl-phenyl)-1H-pyrazolo[3,4- d]pyrimidine 133-135° C. 6

[3-(2-Chloro-5- methanesulfonyl-phenyl)-1H- pyrazolo[3,4-d]pyrimidin-6-yl]-(2,4-difluoro-phenyl)- amine 437 7

1-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- ylamino]-2-methyl-propan-2- ol 525 8

3-[6-(2,4-Difluoro-phenoxy)- 1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methyl- benzenesulfonamide 418 9

3-[6-(2,4-Difluoro-phenoxy)- 1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4,N-dimethyl- benzenesulfonamide 432 10

3-(2-Benzyloxy-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine 509 11

3-[6-(2,4-Difluoro-phenoxy)- 1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4,N,N-trimethyl- benzenesulfonamide 446 12

6-(2,4-Difluoro-phenoxy)-3- (2-ethoxy-5-methanesulfonyl-phenyl)-1H-pyrazolo[3,4- d]pyrimidine 447 13

6-(2,4-Difluoro-phenoxy)-3- [5-methanesulfonyl-2-(2-methoxy-ethoxy)-phenyl]- 1H-pyrazolo[3,4- d]pyrimidine 477 14

3-(5-Chloro-2-ethylsulfanyl- pyridin-4-yl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4- d]pyrimidine 421 15

3-(2-Chloro-5- methylsulfanyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine 406 16

1-{5-Chloro-4-[6-(2,4- difluoro-phenoxy)-1H- pyrazolo[3,4-d]pyrimidin-3-yl]-pyridin-2-ylsulfanyl}- propan-2-ol 451 17

3-{5-Chloro-4-[6-(2,4- difluoro-phenoxy)-1H- pyrazolo[3,4-d]pyrimidin-3-yl]-pyridin-2-ylsulfanyl}- propane-1,2-diol 467 18

3-[6-(2,4-Difluoro-phenoxy)- 1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methoxy-N-methyl- benzenesulfonamide 448 19

3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine-4- carbonitrile 463 20

6-(2,4-Difluoro-phenoxy)-3- (5-methanesulfonyl-2-prop-2-ynyloxy-phenyl)-1H- pyrazolo[3,4-d]pyrimidine 457 21

{2-[6-(2,4-Difluoro- phenoxy)-1H-pyrazolo[3,4- d]pyrimidin-3-yl]-4-methanesulfonyl-phenoxy}- acetonitrile 458 22

4-{2-[6-(2,4-Difluoro- phenoxy)-1H-pyrazolo[3,4- d]pyrimidin-3-yl]-4-methanesulfonyl-phenoxy}- but-2-yn-1-ol 487 23

2-{2-[6-(2,4-Difluoro- phenoxy)-1H-pyrazolo[3,4- d]pyrimidin-3-yl]-4-methanesulfonyl-phenoxy}- ethanol 463 24

2-[6-(2,4-Difluoro-phenoxy)- 1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methanesulfonyl- phenol 419 25

1-[6-(2,4-Difluoro-phenoxy)- 3-(5-methanesulfonyl-2- methyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4- ylamino]-propan-2-ol 490 26

1-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- ylamino]-propan-2-ol 511 27

5-Chloro-4-[6-(2,4-difluoro- phenoxy)-1H-pyrazolo[3,4- b]pyridin-3-yl]-pyridine-2- sulfonic acid methylamide 28

3-(2-Chloro-5-morpholin-4- yl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4- b]pyridine 29

6-(2,4-Difluoro-phenoxy)-3- (5-methanesulfonyl-2-methyl-phenyl)-1H-pyrazolo[4,3- b]pyridine 30

6-(2,4-Difluoro-phenoxy)-3- (5-methanesulfonyl-2-methyl-phenyl)-5-methoxy-1H- pyrazolo[4,3-b]pyridine 31

3-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- yloxy]-propane-1,2-diol >300  32

3-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- yloxy]-propane-1,2-diol 148.2-150.1 33

3-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- ylamino]-propane-1,2-diol 130.0-133.0 34

3-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- ylamino]-propane-1,2-diol 240.7-242.5 35

3-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- ylamino]-pentane-1,5-diol 129.0-136.0 36

2-[3-(2-Chloro-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4- ylamino]-2-methyl-propane- 1,3-diol147.0-151.0 37

6-(2,4-Difluoro-phenoxy)-3- (5-methanesulfonyl-2-trifluoromethoxy-phenyl)-1H- pyrazolo[3,4-d]pyrimidine 38

3-(2-Bromo-5- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine 39

3-(2-Chloro-4- methanesulfonyl-phenyl)-6- (2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine 40

2-[6-(2,4-Difluoro-phenoxy)- 1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methanesulfonyl- phenylamineSynthesis

Compounds of the present invention can be made by a variety of methodsdepicted in the illustrative synthetic reaction schemes shown anddescribed below.

The starting materials and reagents used in preparing these compoundsgenerally are either available from commercial suppliers, such asAldrich Chemical Co., or are prepared by methods known to those skilledin the art following procedures set forth in references such as Fieserand Fieser's Reagents for Organic Synthesis; Wiley & Sons: New York,1991, Volumes 1-15; Rodd's Chemistry of Carbon Compounds, ElsevierScience Publishers, 1989, Volumes 1-5 and Supplementals; and OrganicReactions, Wiley & Sons: New York, 1991, Volumes 1-40. The followingsynthetic reaction schemes are merely illustrative of some methods bywhich the compounds of the present invention can be synthesized, andvarious modifications to these synthetic reaction schemes can be madeand will be suggested to one skilled in the art having referred to thedisclosure contained in this Application.

The starting materials and the intermediates of the synthetic reactionschemes can be isolated and purified if desired using conventionaltechniques, including but not limited to, filtration, distillation,crystallization, chromatography, and the like. Such materials can becharacterized using conventional means, including physical constants andspectral data.

Unless specified to the contrary, the reactions described hereinpreferably are conducted under an inert atmosphere at atmosphericpressure at a reaction temperature range of from about −78° C. to about150° C., more preferably from about 0° C. to about 125° C., and mostpreferably and conveniently at about room (or ambient) temperature,e.g., about 20° C.

One of the specific methods for preparing pyrazolopyrimidine compoundsof the invention is shown in Scheme A below, wherein R is alkyl and p,n, D, E, G, W, X, Y, R^(a), R⁴, R⁵ and R⁶ are as defined herein.

In Step 1 of Scheme A, bisphenoxy compound a is treated with a Grignardreagent b, followed by benzaldehyde (where D, E and G are carbon)compound c, to afford alcohol compound d. Alcohol d is then reduced tothe corresponding keto compound e. Reaction of compound e with hydrazineeffects a ring closure to yield pyrazolo-pyrimidine compound f. Compoundf may then be treated with mild oxidizing reagent such as a peracid, toyield alkylsulfonyl compound g, which is a compound of formula Ia inaccordance with the invention.

In certain embodiments of the invention, benzaldehyde c may be replacedby the corresponding benzoic acid chloride to directly yield ketonecompound e, thus making the oxidation of step 2 unnecessary.

In another variation of Scheme A, R² may be a leaving group such asbenzyl, and instead of sulfur oxidation in step 4, compound f is treatedwith N-chlorosuccinimide under acidic conditions to form abenzenesulfonyl chloride. The benzenesulfonyl chloride may then bereacted with ammonia, alkylamine or dialkylamine to provide compoundswherein R² is amino.

One of skill in the art will understand that certain modifications tothe above schemes are contemplated and within the scope of the presentinvention. For example, where R² is heteroalkyl that requires protection(e.g., aminoalkyl), certain steps will involve the use of protectinggroups for functional groups that are not compatible with particularreaction conditions.

Additional synthetic procedures useful for making compounds of theinvention, including various combinations of aza substitutions for X andY, are disclosed in U.S. patent application Ser. Nos. 11/065,890 and11/067,336 filed on Feb. 25, 2005, the disclosures of which areincorporated herein by reference.

More specific details for producing compounds of the invention aredescribed in the Examples section below.

Pharmaceutical Compositions and Administration

The present invention includes pharmaceutical compositions comprising atleast one compound of the present invention, or an individual isomer,racemic or non-racemic mixture of isomers or a pharmaceuticallyacceptable salt or solvate thereof, together with at least onepharmaceutically acceptable carrier, and optionally other therapeuticand/or prophylactic ingredients.

In general, the compounds of the present invention will be administeredin a therapeutically effective amount by any of the accepted modes ofadministration for agents that serve similar utilities. Suitable dosageranges are typically 1-500 mg daily, preferably 1-100 mg daily, and mostpreferably 1-30 mg daily, depending upon numerous factors such as theseverity of the disease to be treated, the age and relative health ofthe subject, the potency of the compound used, the route and form ofadministration, the indication towards which the administration isdirected, and the preferences and experience of the medical practitionerinvolved. One of ordinary skill in the art of treating such diseaseswill be able, without undue experimentation and in reliance uponpersonal knowledge and the disclosure of this Application, to ascertaina therapeutically effective amount of the compounds of the presentinvention for a given disease.

In general, compounds of the present invention will be administered aspharmaceutical formulations including those suitable for oral (includingbuccal and sub-lingual), rectal, nasal, topical, pulmonary, vaginal, orparenteral (including intramuscular, intraarterial, intrathecal,subcutaneous and intravenous) administration or in a form suitable foradministration by inhalation or insufflation. The preferred manner ofadministration is generally oral using a convenient daily dosage regimenwhich can be adjusted according to the degree of affliction.

A compound or compounds of the present invention, together with one ormore conventional adjuvants, carriers, or diluents, may be placed intothe form of pharmaceutical compositions and unit dosages. Thepharmaceutical compositions and unit dosage forms may be comprised ofconventional ingredients in conventional proportions, with or withoutadditional active compounds or principles, and the unit dosage forms maycontain any suitable effective amount of the active ingredientcommensurate with the intended daily dosage range to be employed. Thepharmaceutical compositions may be employed as solids, such as tabletsor filled capsules, semisolids, powders, sustained release formulations,or liquids such as solutions, suspensions, emulsions, elixirs, or filledcapsules for oral use; or in the form of suppositories for rectal orvaginal administration; or in the form of sterile injectable solutionsfor parenteral use. Formulations containing about one (1) milligram ofactive ingredient or, more broadly, about 0.01 to about one hundred(100) milligrams, per tablet, are accordingly suitable representativeunit dosage forms.

The compounds of the present invention may be formulated in a widevariety of oral administration dosage forms. The pharmaceuticalcompositions and dosage forms may comprise a compound or compounds ofthe present invention or pharmaceutically acceptable salts thereof asthe active component. The pharmaceutically acceptable carriers may beeither solid or liquid. Solid form preparations include powders,tablets, pills, capsules, cachets, suppositories, and dispersiblegranules. A solid carrier may be one or more substances which may alsoact as diluents, flavouring agents, solubilizers, lubricants, suspendingagents, binders, preservatives, tablet disintegrating agents, or anencapsulating material. In powders, the carrier generally is a finelydivided solid which is a mixture with the finely divided activecomponent. In tablets, the active component generally is mixed with thecarrier having the necessary binding capacity in suitable proportionsand compacted in the shape and size desired. The powders and tabletspreferably contain from about one (1) to about seventy (70) percent ofthe active compound. Suitable carriers include but are not limited tomagnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin,dextrin, starch, gelatine, tragacanth, methylcellulose, sodiumcarboxymethylcellulose, a low melting wax, cocoa butter, and the like.The term “preparation” is intended to include the formulation of theactive compound with encapsulating material as carrier, providing acapsule in which the active component, with or without carriers, issurrounded by a carrier, which is in association with it. Similarly,cachets and lozenges are included. Tablets, powders, capsules, pills,cachets, and lozenges may be as solid forms suitable for oraladministration.

Other forms suitable for oral administration include liquid formpreparations including emulsions, syrups, elixirs, aqueous solutions,aqueous suspensions, or solid form preparations which are intended to beconverted shortly before use to liquid form preparations. Emulsions maybe prepared in solutions, for example, in aqueous propylene glycolsolutions or may contain emulsifying agents, for example, such aslecithin, sorbitan monooleate, or acacia. Aqueous solutions can beprepared by dissolving the active component in water and adding suitablecolorants, flavors, stabilizers, and thickening agents. Aqueoussuspensions can be prepared by dispersing the finely divided activecomponent in water with viscous material, such as natural or syntheticgums, resins, methylcellulose, sodium carboxymethylcellulose, and otherwell known suspending agents. Solid form preparations include solutions,suspensions, and emulsions, and may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

The compounds of the present invention may be formulated for parenteraladministration (e.g., by injection, for example bolus injection orcontinuous infusion) and may be presented in unit dose form in ampoules,pre-filled syringes, small volume infusion or in multi-dose containerswith an added preservative. The compositions may take such forms assuspensions, solutions, or emulsions in oily or aqueous vehicles, forexample solutions in aqueous polyethylene glycol. Examples of oily ornonaqueous carriers, diluents, solvents or vehicles include propyleneglycol, polyethylene glycol, vegetable oils (e.g., olive oil), andinjectable organic esters (e.g., ethyl oleate), and may containformulatory agents such as preserving, wetting, emulsifying orsuspending, stabilizing and/or dispersing agents. Alternatively, theactive ingredient may be in powder form, obtained by aseptic isolationof sterile solid or by lyophilization from solution for constitutionbefore use with a suitable vehicle, e.g., sterile, pyrogen-free water.

The compounds of the present invention may be formulated for topicaladministration to the epidermis as ointments, creams or lotions, or as atransdermal patch. Ointments and creams may, for example, be formulatedwith an aqueous or oily base with the addition of suitable thickeningand/or gelling agents. Lotions may be formulated with an aqueous or oilybase and will in general also containing one or more emulsifying agents,stabilizing agents, dispersing agents, suspending agents, thickeningagents, or coloring agents. Formulations suitable for topicaladministration in the mouth include lozenges comprising active agents ina flavored base, usually sucrose and acacia or tragacanth; pastillescomprising the active ingredient in an inert base such as gelatine andglycerine or sucrose and acacia; and mouthwashes comprising the activeingredient in a suitable liquid carrier.

The compounds of the present invention may be formulated foradministration as suppositories. A low melting wax, such as a mixture offatty acid glycerides or cocoa butter is first melted and the activecomponent is dispersed homogeneously, for example, by stirring. Themolten homogeneous mixture is then poured into convenient sized molds,allowed to cool, and to solidify.

The compounds of the present invention may be formulated for vaginaladministration. Pessaries, tampons, creams, gels, pastes, foams orsprays containing in addition to the active ingredient such carriers asare known in the art to be appropriate.

The compounds of the present invention may be formulated for nasaladministration. The solutions or suspensions are applied directly to thenasal cavity by conventional means, for example, with a dropper, pipetteor spray. The formulations may be provided in a single or multidoseform. In the latter case of a dropper or pipette, this may be achievedby the patient administering an appropriate, predetermined volume of thesolution or suspension. In the case of a spray, this may be achieved forexample by means of a metering atomizing spray pump.

The compounds of the present invention may be formulated for aerosoladministration, particularly to the respiratory tract and includingintranasal administration. The compound will generally have a smallparticle size for example of the order of five (5) microns or less. Sucha particle size may be obtained by means known in the art, for exampleby micronization. The active ingredient is provided in a pressurizedpack with a suitable propellant such as a chlorofluorocarbon (CFC), forexample, dichlorodifluoromethane, trichlorofluoromethane, ordichlorotetrafluoroethane, or carbon dioxide or other suitable gas. Theaerosol may conveniently also contain a surfactant such as lecithin. Thedose of drug may be controlled by a metered valve. Alternatively theactive ingredients may be provided in a form of a dry powder, forexample a powder mix of the compound in a suitable powder base such aslactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidine (PVP). The powder carrier will form agel in the nasal cavity. The powder composition may be presented in unitdose form for example in capsules or cartridges of e.g., gelatine orblister packs from which the powder may be administered by means of aninhaler.

When desired, formulations can be prepared with enteric coatings adaptedfor sustained or controlled release administration of the activeingredient. For example, the compounds of the present invention can beformulated in transdermal or subcutaneous drug delivery devices. Thesedelivery systems are advantageous when sustained release of the compoundis necessary and when patient compliance with a treatment regimen iscrucial. Compounds in transdermal delivery systems are frequentlyattached to an skin-adhesive solid support. The compound of interest canalso be combined with a penetration enhancer, e.g., Azone(1-dodecylazacycloheptan-2-one). Sustained release delivery systems areinserted subcutaneously into the subdermal layer by surgery orinjection. The subdermal implants encapsulate the compound in a lipidsoluble membrane, e.g., silicone rubber, or a biodegradable polymer,e.g., polylactic acid.

The pharmaceutical preparations are preferably in unit dosage forms. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

Other suitable pharmaceutical carriers and their formulations aredescribed in Remington: The Science and Practice of Pharmacy 1995,edited by E. W. Martin, Mack Publishing Company, 19th edition, Easton,Pa. Representative pharmaceutical formulations containing a compound ofthe present invention are described in the Examples below.

Utility

Compounds of the invention are useful for, but not limited to, thetreatment of any disorder or disease state in a human, or other mammal,which is exacerbated or caused by excessive or unregulated TNF or p38kinase production by such mammal. Accordingly, the present inventionprovides a method of treating a p38-mediated disease which comprisesadministering an effective amount of a compound of the invention, or apharmaceutically acceptable salt, solvate or prodrug thereof, to asubject or patient in need thereof.

Compounds of the invention are useful for, but not limited to, thetreatment of inflammation in a subject, and for use as antipyretics forthe treatment of fever. Compounds of the invention would be useful totreat arthritis, including but not limited to, rheumatoid arthritis,spondyloarthropathies, gouty arthritis, osteoarthritis, systemic lupuserythematosus and juvenile arthritis, osteoarthritis, gouty arthritisand other arthritic conditions. Such compounds would be useful for thetreatment of pulmonary disorders or lung inflammation, including adultrespiratory distress syndrome, pulmonary sarcoidosis, asthma, silicosis,and chronic pulmonary inflammatory disease. The compounds are alsouseful for the treatment of viral and bacterial infections, includingsepsis, septic shock, gram negative sepsis, malaria, meningitis,cachexia secondary to infection or malignancy, cachexia secondary toacquired immune deficiency syndrome (AIDS), AIDS, ARC (AIDS relatedcomplex), pneumonia, and herpes virus. The compounds are also useful forthe treatment of bone resorption diseases, such as osteoporosis,endotoxic shock, toxic shock syndrome, reperfusion injury, autoimmunedisease including graft vs. host reaction and allograft rejections,cardiovascular diseases including atherosclerosis, thrombosis,congestive heart failure, and cardiac reperfusion injury, renalreperfusion injury, liver disease and nephritis, and myalgias due toinfection.

The compounds are also useful for the treatment of Alzheimer's disease,influenza, multiple sclerosis, cancer, diabetes, systemic lupuserthrematosis (SLE), skin-related conditions such as psoriasis, eczema,burns, dermatitis, keloid formation, and scar tissue formation. Inaddition, compounds of the invention are useful in treatinggastrointestinal conditions such as inflammatory bowel disease, Crohn'sdisease, gastritis, irritable bowel syndrome and ulcerative colitis. Thecompounds are also useful in the treatment of ophthalmic diseases, suchas retinitis, retinopathies, uveitis, ocular photophobia, and of acuteinjury to the eye tissue. The compounds can also be used in treatingangiogenesis, including neoplasia; metastasis; ophthalmologicalconditions such as corneal graft rejection, ocular neovascularization,retinal neovascularization including neovascularization following injuryor infection, diabetic retinopathy, retrolental fibroplasia andneovascular glaucoma; ulcerative diseases such as gastric ulcer;pathological, but non-malignant, conditions such as hemangiomas,including infantile hemangiomas, angiofibroma of the nasopharynx andavascular necrosis of bone; diabetic nephropathy and cardiomyopathy; anddisorders of the female reproductive system such as endometriosis. Thecompounds can further be used for preventing the production ofcyclooxygenase-2 and have analgesic properties. Therefore, Compounds ofFormula I are useful for treatment of pain.

Other uses for Compounds of Formula I include treatment of HCV, severeasthma, psoriasis, chronic obstructive pulmonary disease (COPD), cancer,multiple myeloma, and other diseases that can be treated with ananti-TNF compound.

Besides being useful for human treatment, these compounds are alsouseful for veterinary treatment of companion animals, exotic animals andfarm animals, including mammals, rodents, and the like. More preferredanimals include horses, dogs, and cats.

The present compounds can also be used in co-therapies, partially orcompletely, in place of other conventional anti-inflammatories, such astogether with steroids, cyclooxygenase-2 inhibitors, NSAIDs, DMARDS,immunosuppressive agents, 5-lipoxygenase inhibitors, LTB₄ antagonistsand LTA₄ hydrolase inhibitors.

As used herein, the term “TNF mediated disorder” refers to any and alldisorders and disease states in which TNF plays a role, either bycontrol of TNF itself, or by TNF causing another monokine to bereleased, such as but not limited to IL-1, IL-6 or IL-8. A disease statein which, for instance, IL-1 is a major component, and whose productionor action, is exacerbated or secreted in response to TNF, wouldtherefore be considered a disorder mediated by TNF.

As used herein, the term “p38 mediated disorder” refers to any and alldisorders and disease states in which p38 plays a role, either bycontrol of p38 itself, or by p38 causing another factor to be released,such as but not limited to IL-1, IL-6 or IL-8. A disease state in which,for instance, IL-1 is a major component, and whose production or action,is exacerbated or secreted in response to p38, would therefore beconsidered a disorder mediated by p38.

As TNF-β has close structural homology with TNF-α (also known ascachectin), and since each induces similar biologic responses and bindsto the same cellular receptor, the synthesis of both TNF-α and TNF-β areinhibited by the compounds of the present invention and thus are hereinreferred to collectively as “TNF” unless specifically delineatedotherwise.

EXAMPLES

The following preparations and examples are given to enable thoseskilled in the art to more clearly understand and to practice thepresent invention. They should not be considered as limiting the scopeof the invention, but merely as being illustrative and representativethereof.

Unless otherwise stated, all temperatures including melting points(i.e., MP) are in degrees celsius (° C.). It should be appreciated thatthe reaction which produces the indicated and/or the desired product maynot necessarily result directly from the combination of two reagentswhich were initially added, i.e., there may be one or more intermediateswhich are produced in the mixture which ultimately leads to theformation of the indicated and/or the desired product. The followingabbreviations may be used in the Examples.

Abbreviations

DCM dichloromethane

DMF N,N-dimethylformamide

DMAP 4-dimethylaminopyridine

EtOAc ethyl acetate

EtOH ethanol

gc gas chromatography

HMPA hexamethylphosphoramide

hplc high performance liquid chromatography

mCPBA m-chloroperbenzoic acid

MeCN acetonitrile

MeOH methanol

TEA triethylamine

THF tetrahydrofuran

LDA lithium diisopropylamine

TLC thin layer chromatography

Example 13-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine

The synthetic procedure of Example 1 is outlined in Scheme B.

Step 1 5-Bromo-2,4-bis-(2,4-difluoro-phenoxy)-pyrimidine

Sodium Hydride (35.0 g, 875 mmol) was added to 400 mL dry THF, and themixture was cooled in an ice bath for 30 minutes with stirring.2,4-Difluorophenol (84.0 mL, 879 mmol) was added dropwise to thestirring mixture over a 75 minute period. The ice bath was removed, and5-bromo-2,4-dichloropyrimidine (50.23 g, 220 mL was added. The reactionmixture was heated to 65° C. for 3.5 hours with stirring, then cooled toroom temperature. The reaction mixture was partitioned between 750 mL ofEtOAc and 400 mL water. The organic layer was dried over MgSO₄,filtered, and concentrated under reduced pressure. The residue wasdissolved in 100 mL of hot EtOAc, and 300 mL hexanes was added. Themixture was cooled to room temperature and then refrigerated at 5° C.for 18 hours. The resulting crystalline precipitate was collected byfiltration, washed with cold hexanes/EtOAc (3:1), and dried to give63.66 g (70%) of 5-bromo-2,4-bis-(2,4-difluoro-phenoxy)-pyrimidine. MS(M+H)=416.

Step 2.[2,4-Bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-chloro-5-methylsulfanyl-phenyl)-methanone

To 5-bromo-2,4-bis-(2,4-difluoro-phenoxy)-pyrimidine (3.83 g, 9.23 mmol)in dry THF at 0° C. was added isopropyl magnesium chloride (6.93 mL of2.0 M solution in THF, 13.85 mmol). The resulting mixture was stirred at0° C. for 20 minutes. A solution of 2-chloro-5-methylsulfanyl-benzoylchloride (4.27 g, 19.33 mmol) in 20 mL dry THF was added dropwise, withstirring at 0° C. continued for one hour. The reaction mixture waspartitioned between water and ethyl acetate, and the organic layer wasseparated, washed with saturated brine, dried over MgSO₄, filtered andconcentrated under reduced pressure. The residue was chromatographedthrough silica gel (EtOAc/hexanes 5-25%) to give 3.015 g of[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-chloro-5-methylsulfanyl-phenyl)-methanone,MS (M+H)=521.

Step 3.3-(2-Chloro-5-methylsulfanyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine

To a solution of[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-chloro-5-methylsulfanyl-phenyl)-methanone(3.0 g, 5.76 mmol) in 10 mL of dioxane/EtOH (10:1) was added hydrazine(0.19 mL, 5.76 mmol) The resulting mixture was heated to reflux for 24hours, then cooled to room temperature. The mixture was partitionedbetween water and ethyl acetate, and the organic layer was separated,washed twice with 1N aqueous HCl and once with wtisaturated brine, driedover MgSO₄, filtered and concentrated under reduced pressure. Theresidue was chromatographed through silica gel (EtOAc/hexanes 5-20%) togive 180 mg of3-(2-chloro-5-methylsulfanyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine,MS (M+H)=405.

Step 4.3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine

To a solution of3-(2-chloro-5-methylsulfanyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine(158 mg, 0.39 mmol) in 30 mL THF and 10 mL MeOH was added mCPBA (219 mgof 77% solids). The reaction mixture was stirred for 16 hours at roomtemperature, then partitioned between water and ethyl acetate. Theorganic layer was separated, washed three times with saturated aqueousNaHCO₃ and once with saturated brine, dried over MgSO₄, filtered andconcentrated under reduced pressure. The residue was chromatographedthrough silica gel (EtOAc/hexanes 20-50%) to give 50.9 mg of3-(2-chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine,mp.=251.6-252.2° C., MS (M+H)=437.

Additional compounds made by the above procedure are shown in Table 1.

Example 26-(2,4-Difluoro-phenoxy)-3-(5-methanesulfonyl-2-methyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine

The synthetic procedure of Example 2 is outlined in Scheme C.

Step 1.[2,4-Bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-methyl-5-methylsulfanyl-phenyl)-methanol

5-Bromo-2,4-bis-(2,4-difluoro-phenoxy)-pyrimidine (2.73 g, 6.587 mmol)was dissolved in 20 mL of dry THF, and the solution was cooled to 0° C.Isopropyl magnesium chloride (3.62 mL of 2.0 M solution in THF) wasadded dropwise, and the solution was stirred at 0° C. for thirtyminutes, then cooled to −78° C. A solution of3-methanesulfanyl-6-methyl-benzaldehyde (1.1 g, 6.587 mmol) in 10 mL dryTHF was added dropwise, and the reaction mixture was stirred for onehour during which time the reaction mixture was allowed to warm to roomtemperature. The reaction was quenched by addition of 1N HCl, and theresulting solution was partitioned between water and ethyl acetate, andthe organic layer was separated, dried over MgSO₄, filtered andconcentrated under reduced pressure to yield 2.15 g of crude[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-methyl-5-methylsulfanyl-phenyl)-methanol,which was used in the subsequent step without further purification.

Step 2.[2,4-Bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-methyl-5-methylsulfanyl-phenyl)-methanone

[2,4-Bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-methyl-5-methylsulfanyl-phenyl)-methanol(2.15 g, 4.281 mmol) was dissolved in 40 mL methylene chloride togetherwith 10.0 g of MnO₂, and the reaction mixture was heated to reflux for16 hours. The reaction mixture was cooled and filtered through Celite.The filtrate was concentrated under reduced pressure, and the resultingresidue was recrystallized from methylene chloride/hexanes to yield1.816 g of[2,4-Bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-methyl-5-methylsulfanyl-phenyl)-methanone,MS (M+H)=501.

Step 3.6-(2,4-Difluoro-phenoxy)-3-(2-methyl-5-methylsulfanyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine

Using the procedure of step 2 of Example 1,[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-(2-methyl-5-methylsulfanyl-phenyl)-methanone(1.816 g, 3.67 mmol) was converted to6-(2,4-difluoro-phenoxy)-3-(2-methyl-5-methylsulfanyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine(0.59 g, 1.532 mmol), MS (M+H)=385.

Step 4.6-(2,4-Difluoro-phenoxy)-3-(5-methanesulfonyl-2-methyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine

Using the procedure of step 3 of Example 1,6-(2,4-difluoro-phenoxy)-3-(2-methyl-5-methylsulfanyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine(0.59 g, 1.532 mmol) was converted to 350 mg of6-(2,4-difluoro-phenoxy)-3-(5-methanesulfonyl-2-methyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,MP=133-135° C., MS (M+H)=417.

Additional compounds made by the above procedure are shown in Table 1.

Example 34-Chloro-3-[6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-benzenesulfonamide

The synthetic procedure of Example 1 is outlined in Scheme D.

Step 1(5-Benzylsulfanyl-2-chloro-phenyl)-[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-methanol

Using the procedure of step 1 of Example 2 but replacing3-methanesulfanyl-6-methyl-benzaldehyde with3-benzylsulfanyl-6-methyl-benzaldehyde,5-bromo-2,4-bis-(2,4-difluoro-phenoxy)-pyrimidine (2.0 g, 4.96 mmol) wasconverted to 1.93 g of(5-benzylsulfanyl-2-chloro-phenyl)-[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-methanol.

Step 2(5-Benzylsulfanyl-2-chloro-phenyl)-[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-methanone

Using the procedure of step 2 of Example 2,(5-benzylsulfanyl-2-chloro-phenyl)-[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-methanol(1.93 g, 3.22 mmol) was converted to 2.27 g of(5-benzylsulfanyl-2-chloro-phenyl)-[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-methanone,MS (M+H)=598.

Step 33-(5-Benzylsulfanyl-2-chloro-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine

Using the procedure of step 2 of Example 1,(5-benzylsulfanyl-2-chloro-phenyl)-[2,4-bis-(2,4-difluoro-phenoxy)-pyrimidin-5-yl]-methanone(2.27 g, 3.976 mmol) was converted to 0.47 g of3-(5-benzylsulfanyl-2-chloro-phenyl)-6-(2,4-difluoro-phenoxy)-1H—pyrazolo[3,4-d]pyrimidine,MS (M+H)=482.

Step 44-Chloro-3-[6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-benzenesulfonylchloride

3-(5-Benzylsulfanyl-2-chloro-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine(30 mg, 0.98 mmol), N-chlorosuccinimide (38 mg), NaCl (57 mg), formicacid (1 mL), water (1 mL) were added to methylene chloride (2 mL), andthe resulting mixture was stirred at room temperature for four hours.The reaction was quenched by addition of 1N NaHCO₃ and the resultingmixture was partitioned between water and ethyl acetate. The organiclayer was separated, washed once with saturated brine, dried over MgSO₄,filtered and concentrated under reduced pressure. The residue waspurified by preparative TLC (30% EtOAc in hexanes) to give 34 mg of4-Chloro-3-[6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-benzenesulfonylchloride.

Step 54-Chloro-3-[6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-benzenesulfonamide

4-Chloro-3-[6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-benzenesulfonylchloride (34 mg, 0.735 mmol) was added to a solution of 2M NH₃ in MeOH(1 mL), and the reaction mixture was stirred for 20 minutes. Thereaction mixture was concentrated under reduced pressure, and theresidue was purified by preparative TLC (40% EtOAc in hexanes) to give15 mg of4-chloro-3-[6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-benzenesulfonamide,MS (M+H)=418.

Additional compounds made by the above procedure are shown in Table 1.

Example 4

This example illustrates a p38 (MAP) kinase in vitro assay useful forevaluating the compounds of the invention.

The p38 MAP kinase inhibitory activity of compounds of this invention invitro was determined by measuring the transfer of the γ-phosphate fromγ-³³P-ATP by p-38 kinase to Myelin Basic Protein (MBP), using a minormodification of the method described in Ahn, et al., J. Biol. Chem.266:4220-4227 (1991).

The phosphorylated form of the recombinant p38 MAP kinase wasco-expressed with SEK-1 and MEKK in E. Coli (see, Khokhlatchev, et al.,J. Biol. Chem. 272:11057-11062 (1997)) and then purified by affinitychromatography using a Nickel column.

The phosphorylated p38 MAP kinase was diluted in kinase buffer (20 mM3-(N-morpholino)propanesulfonic acid, pH 7.2, 25 mM β-glycerolphosphate, 5 mM ethylene glycol-bis(beta-aminoethylether)-N,N,N′,N′-tetraacetic acid, 1 mM sodium ortho-vanadate, 1 mMdithiothreitol, 40 mM magnesium chloride). Test compound dissolved inDMSO or only DMSO (control) was added and the samples were incubated for10 min at 30° C. The kinase reaction was initiated by the addition of asubstrate cocktail containing MBP and γ-³³P-ATP. After incubating for anadditional 20 min at 30° C., the reaction was terminated by adding 0.75%phosphoric acid. The phosphorylated MBP was then separated from theresidual γ-³³P-ATP using a phosphocellulose membrane (Millipore,Bedfrod, Mass.) and quantitated using a scintillation counter (Packard,Meriden, Conn.).

Using the above procedure, the compounds of the invention were found tobe inhibitors of p38 MAP kinase. For example,2-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-propane-1,3-diol exhibited a p38 IC₅₀ (uM)of approximately 0.002.

Example 5 In Vitro Assay to Evaluate the Inhibition of LPS-Induced TNF-αProduction in THP1 Cells

This example illustrates an in vitro assay to evaluate the inhibition ofLPS-induced TNF-α production in THP1 cells.

The ability of the compounds of this invention to inhibit the TNF-αrelease is determined using a minor modification of the methodsdescribed in Blifeld, et al. Transplantation, 51:498-503 (1991).

(a) Induction of TNF Biosynthesis:

THP-1 cells are suspended in culture medium [RPMI (Gibco-BRL,Gailthersburg, Md.) containing 15% fetal bovine serum, 0.02 mM2-mercaptoethanol], at a concentration of 2.5×10⁶ cells/mL and thenplated in 96 well plate (0.2 mL aliquots in each well). Test compoundsare dissolved in DMSO and then diluted with the culture medium such thatthe final DMSO concentration is 5%. Twenty five μL aliquots of testsolution or only medium with DMSO (control) are added to each well. Thecells are incubated for 30 min., at 37° C. LPS (Sigma, St. Louis, Mo.)is added to the wells at a final concentration of 0.5 μg/ml, and cellswere incubated for an additional 2 h. At the end of the incubationperiod, culture supernatants are collected and the amount of TNF-αpresent is determined using an ELISA assay as described below.

(b) ELISA Assay:

The amount of human TNF-α present is determined by a specific trappingELISA assay using two anti-TNF-α antibodies (2TNF-H12 and 2TNF-H34)described in Reimund, J. M., et al. GUT. Vol. 39(5), 684-689 (1996).

Polystyrene 96-well plates are coated with 50 μl per well of antibody2TNF-H12 in PBS (10 μg/mL) and incubated in a humidified chamber at 4°C. overnight. The plates are washed with PBS and then blocked with 5%nonfat-dry milk in PBS for 1 hour at room temperature and washed with0.1% BSA (bovine serum albumin) in PBS.

TNF standards are prepared from a stock solution of human recombinantTNF-α (R&D Systems, Minneapolis, Minn.). The concentration of thestandards in the assay began at 10 ng/mL followed by 6 half log serialdilutions.

Twenty five μL aliquots of the above culture supernatants or TNFstandards or only medium (control) are mixed with 25 μL aliquots ofbiotinylated monoclonal antibody 2TNF-H34 (2 μg/mL in PBS containing0.1% BSA) and then added to each well. The samples are incubated for 2hr at room temperature with gentle shaking and then washed 3 times with0.1% BSA in PBS. 50 μl of peroxidase-streptavidin (Zymed, S. SanFrancisco, Calif.) solution containing 0.416 μg/mL ofperoxidase-streptavidin and 0.1% BSA in PBS was added to each well. Thesamples are incubated for an additional 1 hr at room temperature andthen washed 4 times with 0.1% BSA in PBS. Fifty μL of O-phenylenediaminesolution (1 μg/mL O-phenylene-diamine and 0.03% hydrogen peroxide in0.2M citrate buffer pH 4.5) is added to each well and the samples areincubated in the dark for 30 min., at room temperature. Optical densityof the sample and the reference are read at 450 nm and 650 nm,respectively. TNF-α levels are determined from a graph relating theoptical density at 450 nm to the concentration used.

Example 6 In Vitro Assay to Evaluate the Inhibition of LPS-Induced TNF-αProduction in THP1 Cells

This example illustrates an in vivo assay to evaluate the inhibition ofLPS-induced TNF-α production in mice (or rats).

The ability of the compounds of this invention to inhibit the TNF-αrelease, in vivo, is determined using a minor modification of themethods described in described in Zanetti, et. al., J. Immunol.,148:1890 (1992) and Sekut, et. al., J. Lab. Clin. Med., 124:813 (1994).

Female BALB/c mice weighing 18-21 grams (Charles River, Hollister,Calif.) are acclimated for one week. Groups containing 8 mice each aredosed orally either with the test compounds suspended or dissolved in anaqueous vehicle containing 0.9% sodium chloride, 0.5% sodiumcarboxymethyl-cellulose, 0.4% polysorbate 80, 0.9% benzyl alcohol (CMCvehicle) or only vehicle (control group). After 30 min., the mice areinjected intraperitoneally with 20 μg of LPS (Sigma, St. Louis, Mo.).After 1.5 h, the mice are sacrificed by CO₂ inhalation and blood isharvested by cardiocentesis. Blood is clarified by centrifugation at15,600×g for 5 min., and sera are transferred to clean tubes and frozenat −20° C. until analyzed for TNF-α by ELISA assay (BiosourceInternational, Camarillo, Calif.) following the manufacturer's protocol.

Example 7 Adjuvant-Induced Arthritis in Rats

AIA-induced arthritis is evalulated using the procedure of Badger etal., Arthritis & Rheumatism, 43(1) pp 175-183 (2000) AIA is induced by asingle injection of 0.75 mg of parrafin-suspended MycobacteriumButycricum) into male Lewis rats. Hindpaw volume is measured by waterdisplacement on days 15, 20 and 30. A set of control animals is dosedwith tragacanth. Test compounds in 0.5% tragacanth are administeredorally at 3, 10, 30 and 60 mg/kg/day dosages. Indomethacin is used as apositive control. Percentage inhibition of hindpaw edema is calculatedby1−[AIA(treated)/AIA(control)]×100where AIA (treated) and AIA (control) represent the mean paw volume.

Example 8

Formulations

Pharmaceutical preparations for delivery by various routes areformulated as shown in the following Tables. “Active ingredient” or“Active compound” as used in the Tables means one or more of theCompounds of Formula I.

Composition for Oral Administration Ingredient % wt./wt. Activeingredient 20.0% Lactose 79.5% Magnesium stearate  0.5%

The ingredients are mixed and dispensed into capsules containing about100 mg each; one capsule would approximate a total daily dosage.

Composition for Oral Administration Ingredient % wt./wt. Activeingredient 20.0% Magnesium stearate 0.5% Crosscarmellose sodium 2.0%Lactose 76.5% PVP (polyvinylpyrrolidine) 1.0%

The ingredients are combined and granulated using a solvent such asmethanol. The formulation is then dried and formed into tablets(containing about 20 mg of active compound) with an appropriate tabletmachine.

Composition for Oral Administration Ingredient Amount Active compound1.0 g Fumaric acid 0.5 g Sodium chloride 2.0 g Methyl paraben 0.15 gPropyl paraben 0.05 g Granulated sugar 25.5 g Sorbitol (70% solution)12.85 g Veegum K (Vanderbilt Co.) 1.0 g Flavoring 0.035 ml Colorings 0.5mg Distilled water q.s. to 100 ml

The ingredients are mixed to form a suspension for oral administration.

Parenteral Formulation Ingredient % wt./wt. Active ingredient 0.25 gSodium Chloride qs to make isotonic Water for injection 100 ml

The active ingredient is dissolved in a portion of the water forinjection. A sufficient quantity of sodium chloride is then added withstirring to make the solution isotonic. The solution is made up toweight with the remainder of the water for injection, filtered through a0.2 micron membrane filter and packaged under sterile conditions.

Suppository Formulation Ingredient % wt./wt. Active ingredient 1.0%Polyethylene glycol 1000 74.5% Polyethylene glycol 4000 24.5%

The ingredients are melted together and mixed on a steam bath, andpoured into molds containing 2.5 g total weight.

Topical Formulation Ingredients grams Active compound 0.2-2 Span 60 2Tween 60 2 Mineral oil 5 Petrolatum 10 Methyl paraben 0.15 Propylparaben 0.05 BHA (butylated hydroxy anisole) 0.01 Water q.s. 100

All of the ingredients, except water, are combined and heated to about60° C. with stirring. A sufficient quantity of water at about 60° C. isthen added with vigorous stirring to emulsify the ingredients, and waterthen added q.s. about 100 g.

Nasal Spray Formulations

Several aqueous suspensions containing from about 0.025-0.5 percentactive compound are prepared as nasal spray formulations. Theformulations optionally contain inactive ingredients such as, forexample, microcrystalline cellulose, sodium carboxymethylcellulose,dextrose, and the like. Hydrochloric acid may be added to adjust pH. Thenasal spray formulations may be delivered via a nasal spray metered pumptypically delivering about 50-100 microliters of formulation peractuation. A typical dosing schedule is 2-4 sprays every 4-12 hours.

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto.

1. A compound of the formula:

or pharmaceutically acceptable salts thereof, wherein: n is 1; R¹ isoptionally substituted phenyl; R² is —S(O)_(m)R^(a) wherein m is from 0to 2, R^(a) is alkyl, heteroalkyl, amino, alkylamino, dialkylamino,hydroxy or alkoxy, R³ is hydrogen or alkyl; R⁴ is hydrogen orheteroalkyl each R⁵ is independently alkyl, halo, haloalkyl, alkoxy,haloalkoxy, cyano, nitro, hydroxy, amino, heteroalkyl, heterocyclyl,heterocyclylalkyl, hydroxycycloalkyl, cycloalkylalkyl, alkoxyalkyloxy,aryl, heteroaryl, aralkyl, heteroaralkyl, aralkoxy, heteroaralkyoxy,cyanoalkoxy, alkenylalkoxy, alkynylalkoxy, hydroxyalkyloxy,hydroxyalkylalkynylalkoxy, —(CHR^(e))_(s)—C(═O)—R^(f),—(CHR^(e))_(s)—O—C(═O)—R^(f), —(CHR^(e))_(s)—NH—C(═O)—R^(f) or—SO₂—R^(f), wherein R^(e) is hydrogen, alkyl or heteroalkyl; R^(f) isalkyl, hydroxy, amino, alkylamino, heteroalkyl, aryl, aralkyl,heteroaryl, or heterocyclyl; and s is from 0 to 4; X and Y are nitrogen;one or two of D, E and G is nitrogen, or D, E and G are carbon; W is abond, O or NR^(i); or W and R⁴ together form cyano; wherein R^(i) ishydrogen, or alkyl, and A is O.
 2. The compound of claim 1, wherein R³is hydrogen.
 3. The compound of claim 2, wherein R¹ is 2-halophenyl or2,4-dihalophenyl.
 4. The compound of claim 2, wherein W is NR^(i) or Oand R⁴ is heteroalkyl.
 5. The compound of claim 2, wherein W is a bondand R⁴ is hydrogen.
 6. The compound of claim 2, wherein R⁵ is alkyl,alkoxy, halo, alkoxyalkoxy or benzyloxy.
 7. The compound of claim 2,wherein A is O and R¹ is 2,4-dihalophenyl.
 8. The compound of claim 2,wherein m is 0 or 2, and R^(a) is alkyl or hydroxyalkyl.
 9. The compoundof claim 1, wherein said compound is of the formula:

wherein: p is from 0 to 4; each R⁶ is independently halo, alkyl, alkoxy,haloalkyl, haloalkoxy or cyano; and m, n, W, R^(a), R⁴ and R⁵ are asrecited in claim
 1. 10. The compound of claim 9, wherein W is NR^(i) orO and R⁴ is heteroalkyl.
 11. The compound of claim 9, wherein W is abond and R⁴ is hydrogen.
 12. The compound of claim 1, wherein saidcompound is of the formula:

wherein W, R⁴, R⁵ and R^(a) are as recited in claim
 1. 13. The compoundof claim 12, wherein R^(a) is alkyl or hydroxyalkyl, and R⁵ is alkyl,alkoxy, halo, alkoxyalkoxy or benzyloxy.
 14. The compound of claim 13,wherein R^(a) is methyl and R⁵ is methyl, chloro, methoxy, ethoxy,2-(methoxy)-ethoxy or benzyloxy.
 15. A composition comprising: (a) apharmaceutically acceptable excipient; and (b) a compound of claim 1.16. A compound selected from the group consisting of:6-(2,4-Difluoro-phenoxy)-3-(3-methanesulfonyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine;6-(2,4-Difluoro-phenoxy)-3-(5-methanesulfonyl-2-methoxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;2-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-propane-1,3-diol;6-(2,4-Difluoro-phenoxy)-3-(5-methanesulfonyl-2-methyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine[3-(2-Chloro-5-methanesulfonyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl]-(2,4-difluoro-phenyl)-amine;1-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-2-methyl-propan-2-ol;3-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methyl-benzenesulfonamide;3-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-N-dimethyl-benzenesulfonamide;3-(2-Benzyloxy-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine;3-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4,N,N-trimethyl-benzenesulfonamide;6-(2,4-Difluoro-phenoxy)-3-(2-ethoxy-5-methanesulfonyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;6-(2,4-Difluoro-phenoxy)-3[5-methanesulfonyl-2-(2-methoxy-ethoxy)-phenyl]-1H-pyrazolo[3,4-d]pyrimidine;3-(5Chloro-2-ethylsulfanyl-pyridin-4-yl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine;3-(2-Chloro-5-methylsulfanyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine;1-{5-Chloro-4-[6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-pyridin-2-ylsulfanyl}-propan-2-ol;3-{5-Chloro-4-[6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-pyridin-2-ylsulfanyl}-propane-1,2-diol;3-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methoxy-N-methyl-benzenesulfonamide;3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine-4-carbonitrile;6-(2,4-Difluoro-phenoxy)-3-(5-methanesulfonyl-2-prop-2-ynyloxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;{2-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methanesulfonyl-phenoxy}-acetonitrile;4-{2-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methanesulfonyl-phenoxy}-but-2-yn-1-ol;2-{2-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methanesulfonyl-phenoxy}-ethanol;2-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methanesulfonyl-phenol;1-[6-(2,4-Difluoro-phenoxy)-3-(5-methanesulfonyl-2-methyl-phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-propan-2-ol;1-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-propan-2-ol;3-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-yloxy]-propane-1,2-diol;3-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-yloxy]-propane-1,2-diol;3-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-propane-1,2-diol;3-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-propane-1,2-diol;3-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-pentane-1,5-diol;2-[3-(2-Chloro-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino]-2-methyl-propane-1,3-diol;6-(2,4-Difluoro-phenoxy)-3-(5-methanesulfonyl-2-trifluoromethoxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;3-(2-Bromo-5-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine;3-(2-Chloro-4-methanesulfonyl-phenyl)-6-(2,4-difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidine;and2-[6-(2,4-Difluoro-phenoxy)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-4-methanesulfonyl-phenylamine.